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1.
Plant Cell ; 33(4): 1286-1302, 2021 05 31.
Artículo en Inglés | MEDLINE | ID: mdl-33793891

RESUMEN

Photosystem II (PSII) uses solar energy to oxidize water and delivers electrons for life on Earth. The photochemical reaction center of PSII is known to possess two stationary states. In the open state (PSIIO), the absorption of a single photon triggers electron-transfer steps, which convert PSII into the charge-separated closed state (PSIIC). Here, by using steady-state and time-resolved spectroscopic techniques on Spinacia oleracea and Thermosynechococcus vulcanus preparations, we show that additional illumination gradually transforms PSIIC into a light-adapted charge-separated state (PSIIL). The PSIIC-to-PSIIL transition, observed at all temperatures between 80 and 308 K, is responsible for a large part of the variable chlorophyll-a fluorescence (Fv) and is associated with subtle, dark-reversible reorganizations in the core complexes, protein conformational changes at noncryogenic temperatures, and marked variations in the rates of photochemical and photophysical reactions. The build-up of PSIIL requires a series of light-induced events generating rapidly recombining primary radical pairs, spaced by sufficient waiting times between these events-pointing to the roles of local electric-field transients and dielectric relaxation processes. We show that the maximum fluorescence level, Fm, is associated with PSIIL rather than with PSIIC, and thus the Fv/Fm parameter cannot be equated with the quantum efficiency of PSII photochemistry. Our findings resolve the controversies and explain the peculiar features of chlorophyll-a fluorescence kinetics, a tool to monitor the functional activity and the structural-functional plasticity of PSII in different wild-types and mutant organisms and under stress conditions.


Asunto(s)
Complejo de Proteína del Fotosistema II/química , Complejo de Proteína del Fotosistema II/metabolismo , Spinacia oleracea/química , Clorofila/análogos & derivados , Clorofila/química , Diurona/farmacología , Fluorescencia , Luz , Complejo de Proteína del Fotosistema II/efectos de los fármacos , Conformación Proteica , Espectrometría de Fluorescencia , Espectroscopía Infrarroja por Transformada de Fourier , Temperatura , Thermosynechococcus/química
2.
Plant J ; 105(3): 619-638, 2021 02.
Artículo en Inglés | MEDLINE | ID: mdl-33119927

RESUMEN

Chloroplast-to-nucleus retrograde signaling is essential for cell function, acclimation to fluctuating environmental conditions, plant growth and development. The vast majority of chloroplast proteins are nuclear-encoded, and must be imported into the organelle after synthesis in the cytoplasm. This import is essential for the development of fully functional chloroplasts. On the other hand, functional chloroplasts act as sensors of environmental changes and can trigger acclimatory responses that influence nuclear gene expression. Signaling via mobile transcription factors (TFs) has been recently recognized as a way of communication between organelles and the nucleus. In this study, we performed a targeted reverse genetic screen to identify dual-localized TFs involved in chloroplast retrograde signaling during stress responses. We found that CHLOROPLAST IMPORT APPARATUS 2 (CIA2) has a functional plastid transit peptide, and can be located both in chloroplasts and the nucleus. Further, we found that CIA2, along with its homolog CIA2-like (CIL) are involved in the regulation of Arabidopsis responses to UV-AB, high light and heat shock. Finally, our results suggest that both CIA2 and CIL are crucial for chloroplast translation. Our results contribute to a deeper understanding of signaling events in the chloroplast-nucleus cross-talk.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiología , Proteínas de Cloroplastos/metabolismo , Fotosíntesis/fisiología , Estrés Fisiológico/fisiología , Factores de Transcripción/metabolismo , Arabidopsis/efectos de los fármacos , Proteínas de Arabidopsis/genética , Núcleo Celular/metabolismo , Proteínas de Cloroplastos/genética , Cloroplastos/genética , Cloroplastos/metabolismo , Diurona/farmacología , Regulación de la Expresión Génica de las Plantas , Respuesta al Choque Térmico/fisiología , Plantas Modificadas Genéticamente , Transducción de Señal , Estrés Fisiológico/efectos de los fármacos , Factores de Transcripción/genética
3.
Planta ; 256(3): 55, 2022 Aug 06.
Artículo en Inglés | MEDLINE | ID: mdl-35932433

RESUMEN

MAIN CONCLUSION: Guard cell- or mesophyll cell-localized phytochromes do not have a predominant direct light sensory role in red- or blue-light-mediated stomatal opening or far-red-light-mediated stomatal closure of Arabidopsis. The role of phytochromes in blue- and red-light-mediated stomatal opening, and far-red-light- mediated decrease in opening, is still under debate. It is not clear whether reduced stomatal opening in a phytochrome B (phyB) mutant line, is due to phytochrome acting as a direct photosensor or an indirect growth effect. The exact tissue localization of the phytochrome photoreceptor important for stomatal opening is also not known. We studied differences in stomatal opening in an Arabidopsis phyB mutant, and lines showing mesophyll cell-specific or guard cell-specific inactivation of phytochromes. Stomatal conductance (gs) of intact leaves was measured under red, blue, and blue + far-red light. Lines exhibiting guard cell-specific inactivation of phytochrome did not show a change in gs under blue or red light compared to Col-0. phyB consistently exhibited a reduction in gs under both blue and red light. Addition of far-red light did not have a significant impact on the blue- or red-light-mediated stomatal response. Treatment of leaves with DCMU (3-(3,4-dichlorophenyl)-1,1-dimethylurea), a photosynthetic electron transport (PET) inhibitor, eliminated the response to red light in all lines, indicating that stomatal opening under red light is controlled by PET, and not directly by phytochrome. Similar to previous studies, leaves of the phyB mutant line had fewer stomata. Overall, phytochrome does not appear have a predominant direct sensory role in stomatal opening under red or blue light. However, phytochromes likely have an indirect effect on the degree of stomatal opening under light through effects on leaf growth and stomatal development.


Asunto(s)
Proteínas de Arabidopsis/fisiología , Arabidopsis/fisiología , Células del Mesófilo/química , Fitocromo/fisiología , Arabidopsis/citología , Arabidopsis/efectos de la radiación , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/efectos de la radiación , Diurona/farmacología , Transporte de Electrón/fisiología , Herbicidas/farmacología , Luz , Fotosíntesis/fisiología , Fitocromo/genética , Fitocromo B/genética , Fitocromo B/fisiología , Hojas de la Planta/fisiología , Hojas de la Planta/efectos de la radiación , Estomas de Plantas/fisiología , Estomas de Plantas/efectos de la radiación
4.
Photosynth Res ; 153(1-2): 71-82, 2022 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-35389175

RESUMEN

The redox state of the plastoquinone (PQ) pool is a known sensor for retrograde signaling. In this paper, we asked, "does the redox state of the PQ pool modulate the saturation state of thylakoid lipids?" Data from fatty acid composition and mRNA transcript abundance analyses suggest a strong connection between these two aspects in a model marine diatom. Fatty acid profiles of Phaeodactylum tricornutum exhibited specific changes when the redox state of the PQ pool was modulated by light and two chemical inhibitors [3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) or 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB)]. Data from liquid chromatography with tandem mass spectrometry (LC-MS/MS) indicated a ca. 7-20% decrease in the saturation state of all four conserved thylakoid lipids in response to an oxidized PQ pool. The redox signals generated from an oxidized PQ pool in plastids also increased the mRNA transcript abundance of nuclear-encoded C16 fatty acid desaturases (FADs), with peak upregulation on a timescale of 6 to 12 h. The connection between the redox state of the PQ pool and thylakoid lipid saturation suggests a heretofore unrecognized retrograde signaling pathway that couples photosynthetic electron transport and the physical state of thylakoid membrane lipids.


Asunto(s)
Diatomeas , Plastoquinona , Benzoquinonas , Cromatografía Liquida , Diatomeas/metabolismo , Dibromotimoquinona/metabolismo , Diurona/farmacología , Transporte de Electrón , Ácido Graso Desaturasas/análisis , Ácido Graso Desaturasas/metabolismo , Ácidos Grasos/análisis , Luz , Lípidos , Oxidación-Reducción , Plastoquinona/metabolismo , ARN Mensajero/metabolismo , Espectrometría de Masas en Tándem , Tilacoides/metabolismo
5.
Int J Phytoremediation ; 24(7): 778-786, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-34541976

RESUMEN

Diuron herbicide is widely used for weeds control in many kinds of cultivations. It reaches the waterbodies through various fate routes and can adversely threaten non-target organism. The current study was carried out to evaluate the antioxidant activity of Spirulina as feed additive against the toxicity of Diuron concentrations (40 and 80 µg/L) on the edible mollusk Mytilus galloprovincialis during seven days of exposure. Oxidative stress biomarkers were applied on mussel gills and digestive gland, investigating changes in enzymes activities such as catalase (CAT), Glutathione-S-transferase (GST) and Acetylcholinesterase (AChE) and the Malondialdehyde level (MDA). The obtained results show that diuron altered oxidative stress biomarkers in both organs, gills and digestive gland. Performed principle component analysis (PCA) highlighted relationship between biomarkers involved in functional response. Spirulina platensis supplemented diet (1 mg/L), completely ameliorated diuron-induced oxidative stress in mussel tissues. Thus, Spirulina seems to be a promising microalgae and eco-friendly tool helping the health recovery of aquatic animals subjected to environmental stressors.


This study provided recent and new data on the impact of Diuron in marine bivalve and the protective effect of Spirulina against Diuron-induced oxidative stress. The results of our study suggest that the antioxidant potential of Spirulina should be strongly candidate for the phytoremediation of Diuron-aquatic contaminated.


Asunto(s)
Mytilus , Spirulina , Contaminantes Químicos del Agua , Acetilcolinesterasa/metabolismo , Acetilcolinesterasa/farmacología , Animales , Biodegradación Ambiental , Biomarcadores/metabolismo , Diurona/farmacología , Glutatión Transferasa/metabolismo , Glutatión Transferasa/farmacología , Mytilus/metabolismo , Estrés Oxidativo , Spirulina/metabolismo , Contaminantes Químicos del Agua/toxicidad
6.
Int J Mol Sci ; 24(1)2022 Dec 21.
Artículo en Inglés | MEDLINE | ID: mdl-36613535

RESUMEN

Rate-limiting steps in the dark-to-light transition of Photosystem II (PSII) were discovered by measuring the variable chlorophyll-a fluorescence transients elicited by single-turnover saturating flashes (STSFs). It was shown that in diuron-treated samples: (i) the first STSF, despite fully reducing the QA quinone acceptor molecule, generated only an F1(

Asunto(s)
Diurona , Complejo de Proteína del Fotosistema II , Temperatura , Diurona/farmacología , Listas de Espera , Clorofila , Clorofila A , Luz
7.
Physiol Plant ; 172(4): 2217-2225, 2021 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-34050526

RESUMEN

The D1:Val219 residue of Photosystem II in the cyanobacterium Synechocystis sp. PCC 6803 was mutated to alanine or isoleucine, creating the V219A and V219I mutants, respectively. Oxygen evolution was slowed in these mutants, while chlorophyll a fluorescence induction assays indicated slowed electron transfer. As previously observed [Erickson J.M., Rahire, M., Rochaix, J.-D. and Mets. L. (1985) Science, 228, 204-207], the V219I mutant was resistant to 3,4-dichloro-1,1-dimethyl urea (DCMU); however, the V219A strain displayed no DCMU resistance. Additionally, the V219A strain was less sensitive to the addition of formate than the control, while the V219I strain was more sensitive to formate. Both mutant strains were susceptible to photodamage and required protein synthesis for recovery. We hypothesize that the sensitivity to DCMU and the extent of bicarbonate-reversible formate-induced inhibition, as well as the capacity for recovery in cells following photodamage, are influenced by the hydrophobicity of the environment associated with the Val219 residue in D1.


Asunto(s)
Complejo de Proteína del Fotosistema II , Synechocystis , Clorofila , Clorofila A , Diurona/farmacología , Formiatos , Interacciones Hidrofóbicas e Hidrofílicas , Mutación , Complejo de Proteína del Fotosistema II/genética , Plastoquinona , Quinonas , Synechocystis/genética
8.
ScientificWorldJournal ; 2019: 1030236, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31346323

RESUMEN

The unicellular halotolerant cyanobacterium Aphanothece halophytica is a potential dark fermentative producer of molecular hydrogen (H2) that produces very little H2 under illumination. One factor limiting the H2 photoproduction of this cyanobacterium is an inhibition of bidirectional hydrogenase activity by oxygen (O2) obtained from splitting water molecules via photosystem II activity. The present study aimed to investigate the effects of the photosystem II inhibitors carbonyl cyanide m-chlorophenyl hydrazone (CCCP) and 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) on H2 production of A. halophytica under light and dark conditions and on photosynthetic and respiratory activities. The results showed that A. halophytica treated with CCCP and DCMU produced H2 at three to five times the rate of untreated cells, when exposed to light. The highest H2 photoproduction rates, 2.26 ±â€Š0.24 and 3.63 ±â€Š0.26 µmol H2 g-1 dry weight h-1, were found in cells treated with 0.5 µM CCCP and 50 µM DCMU, respectively. Without inhibitor treatment, A. halophytica incubated in the dark showed a significant increase in H2 production compared with cells that were incubated in the light. Only CCCP treatment increased H2 production of A. halophytica during dark incubation, because CCCP functions as an uncoupling agent of oxidative phosphorylation. The highest dark fermentative H2 production rate of 39.50 ±â€Š2.13 µmol H2 g-1 dry weight h-1 was found in cells treated with 0.5 µM CCCP after 2 h of dark incubation. Under illumination, CCCP and DCMU inhibited chlorophyll fluorescence, resulting in a low level of O2, which promoted bidirectional hydrogenase activity in A. halophytica cells. In addition, only CCCP enhanced the respiration rate, further reducing the O2 level. In contrast, DCMU reduced the respiration rate in A. halophytica.


Asunto(s)
Carbonil Cianuro m-Clorofenil Hidrazona/farmacología , Cianobacterias/efectos de los fármacos , Cianobacterias/metabolismo , Diurona/farmacología , Hidrógeno/metabolismo , Complejo de Proteína del Fotosistema II/antagonistas & inhibidores , Respiración de la Célula/efectos de los fármacos , Respiración de la Célula/efectos de la radiación , Clorofila A/metabolismo , Oscuridad , Hidrogenasas/metabolismo , Fotosíntesis/efectos de los fármacos
9.
J Environ Sci Health B ; 54(4): 290-293, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-30633634

RESUMEN

The extensive use of pesticides represents a risk to human health and to the environment. This study aimed to investigate if the exposure to atrazine and diuron, two herbicides widely used in Brazil, could induce changes in the susceptibility profile to aztreonam, colistin and polymyxin B antimicrobials in isolates of P. aeruginosa obtained from soil samples by using the determination of minimum inhibitory concentration (MIC) test. Three isolates had an increase of MIC to aztreonam after exposure to both herbicides and one isolate did not show any MIC change. The MexAB-OprM efflux pump has already been upregulated in these isolates and the herbicides atrazine and diuron did not increase MexAB-OprM overexpression. Therefore, the decrease in aztreonam susceptibility was not directly related to this pump, suggesting that probably other mechanisms should be involved.


Asunto(s)
Antibacterianos/farmacología , Farmacorresistencia Bacteriana/efectos de los fármacos , Herbicidas/farmacología , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/genética , Microbiología del Suelo , Atrazina/farmacología , Aztreonam/farmacología , Proteínas de la Membrana Bacteriana Externa/genética , Brasil , Colistina/farmacología , Diurona/farmacología , Farmacorresistencia Bacteriana/genética , Proteínas de Transporte de Membrana/genética , Pruebas de Sensibilidad Microbiana , Polimixina B/farmacología , Pseudomonas aeruginosa/aislamiento & purificación
10.
Proc Natl Acad Sci U S A ; 112(32): 10044-9, 2015 Aug 11.
Artículo en Inglés | MEDLINE | ID: mdl-26150490

RESUMEN

A fundamental mystery of plant cell biology is the occurrence of "stromules," stroma-filled tubular extensions from plastids (such as chloroplasts) that are universally observed in plants but whose functions are, in effect, completely unknown. One prevalent hypothesis is that stromules exchange signals or metabolites between plastids and other subcellular compartments, and that stromules are induced during stress. Until now, no signaling mechanisms originating within the plastid have been identified that regulate stromule activity, a critical missing link in this hypothesis. Using confocal and superresolution 3D microscopy, we have shown that stromules form in response to light-sensitive redox signals within the chloroplast. Stromule frequency increased during the day or after treatment with chemicals that produce reactive oxygen species specifically in the chloroplast. Silencing expression of the chloroplast NADPH-dependent thioredoxin reductase, a central hub in chloroplast redox signaling pathways, increased chloroplast stromule frequency, whereas silencing expression of nuclear genes related to plastid genome expression and tetrapyrrole biosynthesis had no impact on stromules. Leucoplasts, which are not photosynthetic, also made more stromules in the daytime. Leucoplasts did not respond to the same redox signaling pathway but instead increased stromule formation when exposed to sucrose, a major product of photosynthesis, although sucrose has no impact on chloroplast stromule frequency. Thus, different types of plastids make stromules in response to distinct signals. Finally, isolated chloroplasts could make stromules independently after extraction from the cytoplasm, suggesting that chloroplast-associated factors are sufficient to generate stromules. These discoveries demonstrate that chloroplasts are remarkably autonomous organelles that alter their stromule frequency in reaction to internal signal transduction pathways.


Asunto(s)
Cloroplastos/metabolismo , Nicotiana/metabolismo , Transducción de Señal , Secuencia de Bases , Benzoquinonas/farmacología , Cloroplastos/ultraestructura , Ritmo Circadiano/efectos de los fármacos , Diurona/farmacología , Silenciador del Gen/efectos de los fármacos , Proteínas Fluorescentes Verdes/metabolismo , Modelos Biológicos , Datos de Secuencia Molecular , NADP/metabolismo , Oxidación-Reducción/efectos de los fármacos , Fotosíntesis/efectos de los fármacos , Filogenia , Epidermis de la Planta/efectos de los fármacos , Epidermis de la Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/efectos de los fármacos , Sacarosa/farmacología , Reductasa de Tiorredoxina-Disulfuro/metabolismo , Imagen de Lapso de Tiempo , Nicotiana/efectos de los fármacos
11.
Molecules ; 23(10)2018 Oct 19.
Artículo en Inglés | MEDLINE | ID: mdl-30347671

RESUMEN

Eight alkaloids (1⁻8) were isolated from Ruta graveolens, and their herbicide activities were evaluated through in vitro, semivivo, and in vivo assays. The most relevant results were observed for Compounds 5 and 6⁻8 at 150 µM, which decreased dry biomass by 20% and 23%, respectively. These are significant results since they presented similar values with the positive control, commercial herbicide 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). Based on the performed assays, Compound 5 (graveoline) is classified as an electron-transport inhibitor during the light phase of photosynthesis, as well as a plant-growth regulator. On the other hand, Compounds 6⁻8 inhibited electron and energy transfers, and are also plant-growth inhibitors. These phytotoxic behaviors based on acridone and quinolone alkaloids may serve as a valuable tool in the further development of a new class of herbicides since natural products represent an interesting alternative to replace commercial herbicides, potentially due their low toxicity.


Asunto(s)
Alcaloides/aislamiento & purificación , Metoxaleno/análogos & derivados , Fotosíntesis/efectos de los fármacos , Ruta/química , Alcaloides/química , Alcaloides/farmacología , Biomasa , Diurona/farmacología , Transporte de Electrón , Herbicidas/farmacología , Metoxaleno/química , Metoxaleno/aislamiento & purificación , Metoxaleno/farmacología
12.
Plant Physiol ; 171(1): 530-41, 2016 05.
Artículo en Inglés | MEDLINE | ID: mdl-26956667

RESUMEN

Cyanobacteria have evolved effective adaptive mechanisms to improve photosynthesis and CO2 fixation. The central CO2-fixing machinery is the carboxysome, which is composed of an icosahedral proteinaceous shell encapsulating the key carbon fixation enzyme, Rubisco, in the interior. Controlled biosynthesis and ordered organization of carboxysomes are vital to the CO2-fixing activity of cyanobacterial cells. However, little is known about how carboxysome biosynthesis and spatial positioning are physiologically regulated to adjust to dynamic changes in the environment. Here, we used fluorescence tagging and live-cell confocal fluorescence imaging to explore the biosynthesis and subcellular localization of ß-carboxysomes within a model cyanobacterium, Synechococcus elongatus PCC7942, in response to light variation. We demonstrated that ß-carboxysome biosynthesis is accelerated in response to increasing light intensity, thereby enhancing the carbon fixation activity of the cell. Inhibition of photosynthetic electron flow impairs the accumulation of carboxysomes, indicating a close coordination between ß-carboxysome biogenesis and photosynthetic electron transport. Likewise, the spatial organization of carboxysomes in the cell correlates with the redox state of photosynthetic electron transport chain. This study provides essential knowledge for us to modulate the ß-carboxysome biosynthesis and function in cyanobacteria. In translational terms, the knowledge is instrumental for design and synthetic engineering of functional carboxysomes into higher plants to improve photosynthesis performance and CO2 fixation.


Asunto(s)
Ciclo del Carbono/fisiología , Fotosíntesis/fisiología , Synechococcus/fisiología , Diurona/farmacología , Transporte de Electrón , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Luz , Microscopía Fluorescente/métodos , Organismos Modificados Genéticamente , Ribulosa-Bifosfato Carboxilasa/genética , Ribulosa-Bifosfato Carboxilasa/metabolismo , Synechococcus/efectos de los fármacos
13.
Plant Physiol ; 171(1): 580-9, 2016 05.
Artículo en Inglés | MEDLINE | ID: mdl-27016447

RESUMEN

Plant plasma membrane H(+)-ATPase acts as a primary transporter via proton pumping and regulates diverse physiological responses by controlling secondary solute transport, pH homeostasis, and membrane potential. Phosphorylation of the penultimate threonine and the subsequent binding of 14-3-3 proteins in the carboxyl terminus of the enzyme are required for H(+)-ATPase activation. We showed previously that photosynthesis induces phosphorylation of the penultimate threonine in the nonvascular bryophyte Marchantia polymorpha However, (1) whether this response is conserved in vascular plants and (2) the process by which photosynthesis regulates H(+)-ATPase phosphorylation at the plasma membrane remain unresolved issues. Here, we report that photosynthesis induced the phosphorylation and activation of H(+)-ATPase in Arabidopsis (Arabidopsis thaliana) leaves via sugar accumulation. Light reversibly phosphorylated leaf H(+)-ATPase, and this process was inhibited by pharmacological and genetic suppression of photosynthesis. Immunohistochemical and biochemical analyses indicated that light-induced phosphorylation of H(+)-ATPase occurred autonomously in mesophyll cells. We also show that the phosphorylation status of H(+)-ATPase and photosynthetic sugar accumulation in leaves were positively correlated and that sugar treatment promoted phosphorylation. Furthermore, light-induced phosphorylation of H(+)-ATPase was strongly suppressed in a double mutant defective in ADP-glucose pyrophosphorylase and triose phosphate/phosphate translocator (adg1-1 tpt-2); these mutations strongly inhibited endogenous sugar accumulation. Overall, we show that photosynthesis activated H(+)-ATPase via sugar production in the mesophyll cells of vascular plants. Our work provides new insight into signaling from chloroplasts to the plasma membrane ion transport mechanism.


Asunto(s)
Arabidopsis/metabolismo , Membrana Celular/metabolismo , Fotosíntesis/fisiología , ATPasas de Translocación de Protón/metabolismo , Azúcares/metabolismo , Proteasas ATP-Dependientes/genética , Proteasas ATP-Dependientes/metabolismo , Arabidopsis/efectos de los fármacos , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Diurona/farmacología , Luz , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Proteínas de Transporte de Membrana/genética , Proteínas de Transporte de Membrana/metabolismo , Células del Mesófilo/metabolismo , Mutación , Fosforilación , Hojas de la Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente , ATPasas de Translocación de Protón/genética
14.
Z Naturforsch C J Biosci ; 72(7-8): 315-324, 2017 Jul 14.
Artículo en Inglés | MEDLINE | ID: mdl-28258977

RESUMEN

The effects of short-term treatment with phenylurea (DCMU, isoproturon) and phenol-type (ioxynil) herbicides on the green alga Chlorella kessleri and the cyanobacterium Synechocystis salina with different organizations of photosystem II (PSII) were investigated using pulse amplitude modulated (PAM) chlorophyll fluorescence and photosynthetic oxygen evolution measured by polarographic oxygen electrodes (Clark-type and Joliot-type). The photosynthetic oxygen evolution showed stronger inhibition than the PSII photochemistry. The effects of the studied herbicides on both algal and cyanobacterial cells decreased in the following order: DCMU>isoproturon>ioxynil. Furthermore, we observed that the number of blocked PSII centers increased significantly after DCMU treatment (204-250 times) and slightly after ioxynil treatment (19-35 times) in comparison with the control cells. This study suggests that the herbicides affect not only the acceptor side but also the donor side of PSII by modifications of the Mn cluster of the oxygen-evolving complex. We propose that one of the reasons for the different PSII inhibitions caused by herbicides is their influence, in different extents, on the kinetic parameters of the oxygen-evolving reactions (the initial S0-S1 state distribution, the number of blocked centers SB, the turnover time of Si states, misses and double hits). The relationship between the herbicide-induced inhibition and the changes in the kinetic parameters is discussed.


Asunto(s)
Chlorella/efectos de los fármacos , Herbicidas/farmacología , Complejo de Proteína del Fotosistema II/metabolismo , Synechocystis/efectos de los fármacos , Chlorella/metabolismo , Clorofila/metabolismo , Diurona/farmacología , Relación Dosis-Respuesta a Droga , Transporte de Electrón/efectos de los fármacos , Yodobencenos/farmacología , Nitrilos/farmacología , Oxígeno/metabolismo , Compuestos de Fenilurea/farmacología , Fotosíntesis/efectos de los fármacos , Synechocystis/metabolismo
15.
Plant Cell Physiol ; 57(3): 558-67, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26858287

RESUMEN

Estimation of photosynthesis by Chl fluorescence measurement of cyanobacteria is always problematic due to the interference from respiratory electron transfer and from phycocyanin fluorescence. The interference from respiratory electron transfer could be avoided by the use of DCMU or background illumination by blue light, which oxidizes the plastoquinone pool that tends to be reduced by respiration. On the other hand, the precise estimation of photosynthesis in cells with a different phycobilisome content by Chl fluorescence measurement is difficult. By subtracting the basal fluorescence due to the phycobilisome and PSI, it becomes possible to estimate the precise maximum quantum yield of PSII in cyanobacteria. Estimated basal fluorescence accounted for 60% of the minimum fluorescence, resulting in a large difference between the 'apparent' yield and 'true' yield under high phycocyanin conditions. The calculated value of the 'true' maximum quantum yield of PSII was around 0.8, which was similar to the value observed in land plants. The results suggest that the cause of the apparent low yield reported in cyanobacteria is mainly ascribed to the interference from phycocyanin fluorescence. We also found that the 'true' maximum quantum yield of PSII decreased under nitrogen-deficient conditions, suggesting the impairment of the PSII reaction center, while the 'apparent' maximum quantum yield showed a marginal change under the same conditions. Due to the high contribution of phycocyanin fluorescence in cyanobacteria, it is essential to eliminate the influence of the change in phycocyanin content on Chl fluorescence measurement and to evaluate the 'true' photosynthetic condition.


Asunto(s)
Clorofila/metabolismo , Luz , Nitrógeno/deficiencia , Complejo de Proteína del Fotosistema II/metabolismo , Synechocystis/metabolismo , Diurona/farmacología , Fluorescencia , Cinética , Oxidación-Reducción/efectos de los fármacos , Fotosíntesis/efectos de los fármacos , Complejo de Proteína del Fotosistema I/metabolismo , Ficocianina/metabolismo , Synechocystis/efectos de los fármacos
16.
Plant Cell Physiol ; 57(7): 1495-1509, 2016 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-27273581

RESUMEN

Natural capacity has evolved in higher plants to absorb and harness excessive light energy. In basic models, the majority of absorbed photon energy is radiated back as fluorescence and heat. For years the proton sensor protein PsbS was considered to play a critical role in non-photochemical quenching (NPQ) of light absorbed by PSII antennae and in its dissipation as heat. However, the significance of PsbS in regulating heat emission from a whole leaf has never been verified before by direct measurement of foliar temperature under changing light intensity. To test its validity, we here investigated the foliar temperature changes on increasing and decreasing light intensity conditions (foliar temperature dynamics) using a high resolution thermal camera and a powerful adjustable light-emitting diode (LED) light source. First, we showed that light-dependent foliar temperature dynamics is correlated with Chl content in leaves of various plant species. Secondly, we compared the foliar temperature dynamics in Arabidopsis thaliana wild type, the PsbS null mutant npq4-1 and a PsbS-overexpressing transgenic line under different transpiration conditions with or without a photosynthesis inhibitor. We found no direct correlations between the NPQ level and the foliar temperature dynamics. Rather, differences in foliar temperature dynamics are primarily affected by stomatal aperture, and rapid foliar temperature increase during irradiation depends on the water status of the leaf. We conclude that PsbS is not directly involved in regulation of foliar temperature dynamics during excessive light energy episodes.


Asunto(s)
Proteínas de Plantas/metabolismo , Estomas de Plantas/fisiología , Plantas/metabolismo , Temperatura , Diurona/farmacología , Luz , Modelos Lineales , Modelos Biológicos , Especificidad de Órganos/efectos de los fármacos , Especificidad de Órganos/efectos de la radiación , Fotosíntesis/efectos de los fármacos , Fotosíntesis/efectos de la radiación , Estomas de Plantas/efectos de los fármacos , Estomas de Plantas/efectos de la radiación , Transpiración de Plantas/efectos de los fármacos , Transpiración de Plantas/efectos de la radiación , Plantas/efectos de los fármacos , Plantas/efectos de la radiación
17.
Planta ; 244(6): 1303-1313, 2016 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-27541495

RESUMEN

MAIN CONCLUSION: Based on the effects of inorganic salts on chloroplast Fe uptake, the presence of a voltage-dependent step is proposed to play a role in Fe uptake through the outer envelope. Although iron (Fe) plays a crucial role in chloroplast physiology, only few pieces of information are available on the mechanisms of chloroplast Fe acquisition. Here, the effect of inorganic salts on the Fe uptake of intact chloroplasts was tested, assessing Fe and transition metal uptake using bathophenantroline-based spectrophotometric detection and plasma emission-coupled mass spectrometry, respectively. The microenvironment of Fe was studied by Mössbauer spectroscopy. Transition metal cations (Cd2+, Zn2+, and Mn2+) enhanced, whereas oxoanions (NO3-, SO42-, and BO33-) reduced the chloroplast Fe uptake. The effect was insensitive to diuron (DCMU), an inhibitor of chloroplast inner envelope-associated Fe uptake. The inorganic salts affected neither Fe forms in the uptake assay buffer nor those incorporated into the chloroplasts. The significantly lower Zn and Mn uptake compared to that of Fe indicates that different mechanisms/transporters are involved in their acquisition. The enhancing effect of transition metals on chloroplast Fe uptake is likely related to outer envelope-associated processes, since divalent metal cations are known to inhibit Fe2+ transport across the inner envelope. Thus, a voltage-dependent step is proposed to play a role in Fe uptake through the chloroplast outer envelope on the basis of the contrasting effects of transition metal cations and oxoaninons.


Asunto(s)
Transporte Biológico Activo/fisiología , Cloroplastos/metabolismo , Hierro/metabolismo , Beta vulgaris/metabolismo , Beta vulgaris/fisiología , Transporte Biológico Activo/efectos de los fármacos , Cadmio/metabolismo , Cloroplastos/efectos de los fármacos , Cloroplastos/fisiología , Diurona/farmacología , Herbicidas/farmacología , Manganeso/metabolismo , Espectroscopía de Mossbauer , Zinc/metabolismo
18.
Plant Physiol ; 169(1): 717-36, 2015 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-26169679

RESUMEN

Chlamydomonas sp. UWO 241 (UWO 241) is a psychrophilic green alga isolated from Antarctica. A unique characteristic of this algal strain is its inability to undergo state transitions coupled with the absence of photosystem II (PSII) light-harvesting complex protein phosphorylation. We show that UWO 241 preferentially phosphorylates specific polypeptides associated with an approximately 1,000-kD pigment-protein supercomplex that contains components of both photosystem I (PSI) and the cytochrome b6/f (Cyt b6/f) complex. Liquid chromatography nano-tandem mass spectrometry was used to identify three major phosphorylated proteins associated with this PSI-Cyt b6/f supercomplex, two 17-kD PSII subunit P-like proteins and a 70-kD ATP-dependent zinc metalloprotease, FtsH. The PSII subunit P-like protein sequence exhibited 70.6% similarity to the authentic PSII subunit P protein associated with the oxygen-evolving complex of PSII in Chlamydomonas reinhardtii. Tyrosine-146 was identified as a unique phosphorylation site on the UWO 241 PSII subunit P-like polypeptide. Assessment of PSI cyclic electron transport by in vivo P700 photooxidation and the dark relaxation kinetics of P700(+) indicated that UWO 241 exhibited PSI cyclic electron transport rates that were 3 times faster and more sensitive to antimycin A than the mesophile control, Chlamydomonas raudensis SAG 49.72. The stability of the PSI-Cyt b6/f supercomplex was dependent upon the phosphorylation status of the PsbP-like protein and the zinc metalloprotease FtsH as well as the presence of high salt. We suggest that adaptation of UWO 241 to its unique low-temperature and high-salt environment favors the phosphorylation of a PSI-Cyt b6/f supercomplex to regulate PSI cyclic electron transport rather than the regulation of state transitions through the phosphorylation of PSII light-harvesting complex proteins.


Asunto(s)
Chlamydomonas/metabolismo , Complejo de Citocromo b6f/metabolismo , Complejos Multiproteicos/metabolismo , Complejo de Proteína del Fotosistema I/metabolismo , Secuencia de Aminoácidos , Antimicina A/farmacología , Fraccionamiento Químico , Chlamydomonas/efectos de los fármacos , Clorofila/metabolismo , Clorofila A , Diurona/farmacología , Transporte de Electrón/efectos de los fármacos , Electroforesis en Gel Bidimensional , Immunoblotting , Datos de Secuencia Molecular , Complejos Multiproteicos/aislamiento & purificación , Fosforilación/efectos de los fármacos , Estabilidad Proteica/efectos de los fármacos , Alineación de Secuencia , Cloruro de Sodio/farmacología , Tilacoides/efectos de los fármacos , Tilacoides/metabolismo
19.
Photosynth Res ; 129(1): 85-92, 2016 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-27236700

RESUMEN

Although it has been believed that wild-type plants are capable of protecting photosystem I (PSI) under high light, our previous study indicates that PSI is sensitive to high light in the shade-established tree species Psychotria rubra. However, the underlying physiological mechanisms are unclear. In this study, we examined the roles of electron transfer from PSII to PSI and PSI redox state in PSI photoinhibition in P. rubra by treatments with lincomycin (Lin), diuron (DCMU), and methyl viologen (MV). After exposure to 2000 µmol photons m(-2) s(-1) for 2 h, PSI activity decreased by 35, 29, 3, and 49 % in samples treated with H2O, Lin, DCMU, and MV, respectively. Meanwhile, the MV-treated samples showed higher P700 oxidation ratio than the H2O-treated samples, suggesting the PSI photoinhibition under high light was accompanied by high levels of P700 oxidation ratio. PSI photoinhibition was alleviated in the DCMU-treated samples but was accelerated in the MV-treated samples, suggesting that PSI photoinhibition in P. rubra was mainly controlled by electron transfer from PSII to PSI. Taking together, PSI photoinhibition is more related to electron transfer from PSII to PSI rather than PSI redox state in P. rubra, which is different from the mechanisms of PSI photoinhibition in Arabidopsis thaliana and cucumber.


Asunto(s)
Complejo de Proteína del Fotosistema I/antagonistas & inhibidores , Complejo de Proteína del Fotosistema II/antagonistas & inhibidores , Psychotria/fisiología , Diurona/farmacología , Transporte de Electrón , Lincomicina/farmacología , Oxidación-Reducción , Paraquat/farmacología , Fotosíntesis/fisiología , Psychotria/efectos de la radiación , Árboles
20.
Photosynth Res ; 128(1): 15-33, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-26482588

RESUMEN

Lichens are poikilohydric symbiotic organisms that can survive in the absence of water. Photosynthesis must be highly regulated in these organisms, which live under continuous desiccation-rehydration cycles, to avoid photooxidative damage. Analysis of chlorophyll a fluorescence induction curves in the lichen microalgae of the Trebouxiophyceae Asterochloris erici and in Trebouxia jamesii (TR1) and Trebouxia sp. (TR9) phycobionts, isolated from the lichen Ramalina farinacea, shows differences with higher plants. In the presence of the photosynthetic electron transport inhibitor DCMU, the kinetics of Q(A) reduction is related to variable fluorescence by a sigmoidal function that approaches a horizontal asymptote. An excellent fit to these curves was obtained by applying a model based on the following assumptions: (1) after closure, the reaction centers (RCs) can be converted into "energy sink" centers (sRCs); (2) the probability of energy leaving the sRCs is very low or zero and (3) energy is not transferred from the antenna of PSII units with sRCs to other PSII units. The formation of sRCs units is also induced by repetitive light saturating pulses or at the transition from dark to light and probably requires the accumulation of reduced Q(A), as well as structural changes in the reaction centers of PSII. This type of energy sink would provide a very efficient way to protect symbiotic microalgae against abrupt changes in light intensity.


Asunto(s)
Chlorophyta/metabolismo , Líquenes/fisiología , Microalgas/metabolismo , Complejo de Proteína del Fotosistema II/metabolismo , Ascomicetos/fisiología , Clorofila/metabolismo , Clorofila A , Chlorophyta/fisiología , Diurona/farmacología , Líquenes/metabolismo , Luz , Microalgas/efectos de los fármacos , Simbiosis/fisiología
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